International Journal of Mass Spectrometry 413 (2017) 168–175

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International Journal of Mass Spectrometry

journal homepage: www.elsevier.com/locate/ijms

Parallel accumulation for 100% duty cycle trapped ion mobility-mass

spectrometry
Joshua A. Silveira a , Mark E. Ridgeway a , Frank H. Laukien a , Matthias Mann b ,
Melvin A. Park a,∗
a
Bruker Daltonics Inc., Billerica, MA 01821, United States
b
Proteomics and Signal Transduction, Max-Planck-Institute of Biochemistry, Am Klopferspitz 18, 82152 Martinsried, Germany

a r t i c l e i n f o a b s t r a c t

Article history: A new trapped ion mobility spectrometry (TIMS) apparatus has been developed and evaluated. Building
Received 5 February 2016 on the original prototype design, the new spectrometer includes an upstream “ion accumulation trap”.
Accepted 7 March 2016 Inclusion of the new trap allows for storage of a population of ions during the analysis of a previously
Available online 14 March 2016
stored population of ions. By accumulating and analyzing ions in parallel, rather than in series, a duty
cycle of 100% can be achieved. The new design retains the flexibility of the original design—the ability to
Keywords:
exchange mobility range, resolution, and speed/duty cycle—while in all instances, maintaining superior
TIMS
ion utilization efficiency. When operating at 100% duty cycle, the spectrometer is shown to trap a broad
Ion mobility-mass spectrometry
Duty cycle
range of ions (m/z 622–2722) with an average trapping efficiency of ∼70%. Trapping efficiency was found
Parallel accumulation to be constant over a range of accumulation/analysis times from 20 up to 85 ms where an average resolv-
Charge capacity ing power of 60 was achieved. Compared to transmission mode (MS-only) operation, the combination
of high mobility resolving power, high ion utilization efficiency, and relatively fast scan times achieved
with TIMS is shown to result in an improvement in mass spectral peak intensities of more than an order
of magnitude.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction end, while ion-neutral collisions give rise to an opposing steady-

Relative to mass spectrometry (MS), the field of ion mobil-
ity spectrometry (IMS) has been slow to mature. Early reports of
hybrid IMS-MS instruments used to characterize the physical prop-
erties of gaseous ions began to emerge in the 1960s [1,2]. IMS was
introduced in the 1970s as a rapid separations technique—“plasma
chromatography”—and has been widely used thereafter as a stan-
dalone chemical sensor [3]. Still, only in the last couple of decades
have a number of technological advances and commercial sys-
tems extended the use of IMS-MS to a broad range of applications,
including detection of explosives and chemical warfare agents [4,5],
characterization of polymers [6,7], cluster assemblies [8–12], and
biomolecular structures [13–25], as well as high throughput sepa-
ration of complex mixtures [26–32].
Dispersive IMS is conventionally carried out in a drift tube that
contains an inert buffer gas. During IMS analysis, a weak electric
field guides ions injected at one end of the tube toward the opposite
∗ Corresponding author. Tel.: +1 978 663 3600.
E-mail address: mel.park@bruker.com (M.A. Park).
state drag force. Separation results from a disparate number of
collisions experienced by ions of different mobilities (K). During
drift, spatial broadening of the ion swarm stems from thermal dif-
fusion and Coulomb repulsion which principally limit the resolving
power and the number of ions that can be analyzed.
Modern dispersive IMS systems are routinely coupled to time-
of-flight (TOF) MS, owing to the relative speed of the two
techniques. That is, IMS analysis typically occurs on a timescale
between 10 and 100 ms. Assuming a resolving power of 50, IMS
peaks will have a temporal width between 0.2 and 2 ms. In contrast,
TOF MS analysis only requires between 0.1 and 0.2 ms ensuring
that 1–20 mass spectra can be acquired during elution of an IMS
peak. Thus, hybridization of IMS with TOF MS provides a means for
efficiently handling the ions to produce IMS-MS spectra.
In the 1990s, a significant improvement in IMS-MS sensitivity
was achieved when radio-frequency (RF) ion funnels were inte-
grated into hybrid IMS-TOF MS instruments [33,34]. Developed in
response to then-emerging “soft” ionization techniques (i.e., ESI
and MALDI) [35,36], ion funnels were shown to collect, confine,
and efficiently transfer ions through differentially pumped regions
of a mass spectrometer. Incorporating ion funnels at the front and

http://dx.doi.org/10.1016/j.ijms.2016.03.004
1387-3806/© 2016 Elsevier B.V. All rights reserved.
 J.A. Silveira et al. / International Journal of Mass Spectrometry 413 (2017) 168–175
back of a drift tube has also improved the efficiency of ion injection
as well as the collection of the radially diffuse ion swarm [37].
A subsequent ion funnel design featured a dedicated trapping
region for enhanced utilization efficiency when used to pulse ions
into a drift tube [38]. Still, drift tube IMS demands that the initial
temporal width of the ion packet be very small with respect to the
drift time (∼1%) to achieve high resolving power (R ≥ 50). For typical
drift times of ∼50 ms, this limits injection times to a few tenths of
milliseconds and places a major constraint on the instrument duty
cycle. Though proof-of-principle temporal multiplexing strategies
have been shown to increase the ion throughput [39], challenges
due to peak aliasing and the finite duty cycle limit of ∼50% remain.
In addition, temporal multiplexing strategies eliminate the ability
to perform LC-IMS-MS and LC-IMS-MS/MS due to the need to recon-
struct the IMS-MS dataset over a complete multiplexing sequence.
A more recent report described the use of helium (as an alternative
to nitrogen) in the trapping region [40]. Owing to the greater mobil-
ity of ions in a lighter gas, an increase in the charge density of the
injected ion packet was observed, resulting in a sensitivity improve-
ment of more than an order of magnitude. Though more complete
ejection of the accumulated ions was reported when using helium,
the IMS duty cycle (1.6–33%) was still far from 100% since the capac-
ity of the ion funnel/trap was not sufficient to store ions for the
entire duration of the 60 ms drift time.
Introduced in 2011 by Park and coworkers, trapped ion mobility
spectrometry (TIMS) was developed as a highly versatile alterna-
tive to conventional drift tube IMS [41–43]. The general principles
of TIMS [44] and a theory [45,46] have been recently reported. In
TIMS, ions are accumulated and stored according to their mobil-
ity in a manner analogous to the condensed phase electrophoretic
technique termed “electric field gradient focusing”. However, TIMS
also contains an elution step wherein ions are released and sepa-
rated along an electric field gradient (EFG) plateau. The complete
analysis sequence occurs inside an ion funnel located in the first
vacuum stage of the instrument. Among other variables, the resolv-
ing power has been shown to be dependent on the scan rate of the
electric field (ˇ) during the elution step, as well as the drag force act-
ing on the ions. In agreement with theory, experimental conditions
that maximize the work done on ions—longer scans that maxi-
mize the “effective path length” and/or elevated pressures/higher
gas velocities that increase the drag force—have yielded resolving
power up to ∼230 for singly charged ions, ∼270 for doubly charged
peptide ions [47] and ∼300 for multiply charged protein ions [48].
Though exceptional performance has been demonstrated, thus
far, TIMS has operated on the basis of an experimental sequence
wherein ions are sequentially accumulated and analyzed.
In the present work, we report a new spectrometer and
operational scheme that significantly improves the overall TIMS-
TOF MS instrument performance. The new design features a
tunnel—roughly twice the length of the original design—that con-
tains an ion accumulation trap upstream of the analysis region.
Spatially decoupling the accumulation from the analysis region
allows the events to occur simultaneously, thus enabling TIMS
operation at 100% duty cycle. As demonstrated herein, operating
the new spectrometer in this manner yields high TIMS resolving
power, high ion utilization efficiencies, and marked improvements
in the TOF signal intensities, making this approach particularly well
suited for high throughput analyses and other applications requir-
ing high sensitivity.
2. Experimental
A schematic representation of the first-generation spectrome-
ter, its operation, and the coordinate system referred to herein is
shown in Fig. 1a. The TIMS funnel was incorporated into the first
169
vacuum stage of a prototype impact ESI-QqTOF (Bruker Daltonics,
Billerica, MA) mass spectrometer. As depicted in Fig. 1a, the appa-
ratus is comprised of a set of electrodes that form three regions:
the entrance funnel, the TIMS tunnel, and the exit funnel. Ions and
N2 (g) are introduced to the first vacuum stage through the capil-
lary. During operation, ions are deflected into the entrance funnel
while gas may be pumped through a port positioned opposite to
the capillary exit. Alternatively, during TIMS operation, gas flow
through the port is partially or completely restricted such that
gas is diverted through the TIMS tunnel and is pumped through
a secondary port located in the exit funnel region. Specific details
of the first-generation TIMS instrumentation can be found in the
literature [45,46].
The second-generation TIMS funnel, featuring a 96 mm-long
tunnel, was tested on a prototype microTOF (Bruker Daltonics, Bil-
lerica, MA) mass spectrometer. In this design, the entrance and exit
funnel regions are identical to the original design; however, the
tunnel length is increased by roughly a factor of two over that of
the first-generation prototype (46 mm). Doubling the length of the
tunnel allows for the inclusion of a new “ion accumulation trap”
upstream of the analysis region. The accumulation trap enables
a mode of operation wherein incoming ions from the entrance
funnel can be accumulated, while simultaneously, a previously
accumulated population of ions is mobility-analyzed in the “anal-
ysis region”. Inasmuch as ions are continuously accumulated, this
mode of operation eliminates the need to block ions at the deflec-
tion plate and, by definition, allows for a duty cycle of 100%.
The second-generation TIMS funnel has three potential modes of
operation: (1) MS-only/transmission mode wherein the DC fields
and gas flow continuously guide the continuous ion beam to the
detector, (2) serial TIMS operation wherein ions are sequentially
accumulated and analyzed, and (3) parallel accumulation TIMS
operation wherein ion accumulation and analysis occur in parallel.
In MS-only mode, ions are simply transmitted from the entrance
funnel to the exit funnel and subsequently, downstream to the
mass spectrometer without mobility analysis. In serial TIMS mode,
ions are accumulated and mobility-analyzed sequentially, as previ-
ously described [45]. However, with the second-generation design,
one may take advantage of the additional storage capacity afforded
by the longer tunnel [49]. Finally, in parallel accumulation mode,
one may achieve a 100% duty cycle, as outlined above. How-
ever, by using the deflection plate to block ions for part of the
experimental cycle, it is also possible to employ longer analysis
times (while maintaining relatively short accumulation times to
avoid charge capacity issues) when high resolution IMS analysis is
desired.
The entrance and exit funnel regions of the second-generation
spectrometer were pumped using a Roots-type multistage dry vac-
uum pump (∼600 m3 /h, Alcatel, 601B Roots Blower) backed by an
Ebara SA-20 mechanical pump that yielded a pressure of 3.1 and
0.9 mbar in each respective region. The pressure difference gener-
ated across the tunnel has been shown to yield gas velocities on
the order of ∼150 m/s [46]. Ions exiting the TIMS funnel are guided
into the TOF accelerator operated at ∼11 Hz. Both instruments were
equipped with an electrospray ionization (ESI) source operated
with a 5 L/min N2 (g) flow rate. For all experiments, the shape of
the EFG rising edge with respect to position followed a square root
function that has been previously shown to yield equivalent axial
storage space across the mobility range [49].
To characterize the instrument performance, a 1 pmol/␮L tryp-
tic digest of bovine serum albumin (BSA, Protea Biosciences,
Morgantown, WV) was directly infused at ∼150 ␮L/min from a
50:50 mixture of water:acetonitrile containing 0.1% formic acid.
In addition, low concentration tuning mix (Agilent Technologies,
Santa Clara, CA) was used to determine the resolving power by Eq.
(1), efficiency by Eq. (2), and signal enhancement factor by Eq. (3)
170 J.A. Silveira et al. / International Journal of Mass Spectrometry 413 (2017) 168–175

Fig. 1. Comparison of the original 46 mm-long tunnel and serial analysis scheme (a) with the 96 mm-long TIMS tunnel and parallel accumulation scheme (b). In (c), the duty
cycle is plotted as a function of accumulation time for serial (purple line) and parallel accumulation schemes (dotted blue line). In the serial mode of operation, the sum of
the elution and trap steps were assumed to be 50 ms. Note that the transfer step in (b) is very small (<1%) compared to the accumulation/elution step. Furthermore, ions
entering the accumulation trap during the transfer step are immediately transmitted to the analyzer, thus allowing for a duty cycle which is truly 100%.

for TIMS experiments where a broad mass range (m/z 622–2722)
was analyzed:
R = K/K
SignalTIMS 1 seconds. During the elution step, the magnitude of the EFG profile
Efficiency = 100% · · (2)
SignalTrans Duty Cycle
ITIMS
Enhancement factor =
ITrans
In Eqs. (1)–(3), R is the resolving power and K is the ion mobility
coefficient. ITIMS and ITrans are taken to be the maximum intensity
observed for a species during TIMS and the constant intensity of
a species contained in the continuous beam in MS-only experi-
ments, respectively. It is important to recognize that efficiency, as
defined herein, is a measure of the complete system (particularly
with respect to tuning of the entrance funnel, TIMS tunnel, and exit
funnel) when operating in parallel accumulation TIMS mode versus
the system as a whole in MS-only mode. SignalTIMS , used for cal-
culating the efficiency, is taken to be the area under an IMS-MS
peak whereas SignalTrans is the chromatographic area observed in
MS-only mode during the time required for a TIMS experiment.
3. Results and discussion
3.1. Ion storage in TIMS
As depicted in Fig. 1, a feature of TIMS is that the trapping posi-
tion for a particular ion is related to the ion mobility coefficient. The
drag force exerted by the gas flow on an ion in the TIMS tunnel is a
simple function of the ion’s mobility, qvg /K. Also, the counteracting
force from the DC electric field is a function of the axial position
(proportional to qz1/2 ) as implied by the plot in Fig. 1. Accordingly,
ions of low K are dragged by the gas to an axial position further into
the tunnel where the strength of the electric field on the rising edge
is greater. Conversely, ions of higher K are trapped at a position near
the tunnel entrance where the strength of the electric field on the
rising edge is relatively weaker.
A serial analysis sequence (see Fig. 1a) is comprised of accu-
mulation, blocking, and elution steps. The second blocking step
involves preventing additional ions from entering the entrance
funnel/tunnel regions by pulsing the potential of the deflector lens
to an attractive potential. During this time, trapped ions acquire
an equilibrium position along the rising edge and are held for a
(1)
user-defined time period, typically, on the order of a few milli-
is decreased from an initial value at a fixed scan rate such that
ions of progressively higher K elute from the analyzer. Given that
scanning the EFG at a slower rate is known to improve the resolv-
(3)
ing power, the ability to vary the time during the accumulation
and scanning steps makes it possible to trade speed/duty cycle for
resolving power.
3.2. Theoretical charge capacity estimation
It is, of course, natural that any device used to store ions should
have a limited charge capacity. Therefore, part of our effort in
understanding TIMS must be to determine an approximate charge
capacity and the consequences of exceeding this finite limit. In
considering the operation of Paul traps, one may consider the stor-
age capacity (i.e., the maximum number of charges that may be
confined in the physical dimensions of the trap) and the analyt-
ical capacity (i.e., the maximum number of charges that may be
confined without substantially degrading analytical performance).
Furthermore, it is known that in Paul traps, the distribution of
charges with respect to m/z has an influence on relative perfor-
mance as a function of mass. That is, if the ion density is relatively
high in a narrow m/z range, the performance of the analyzer in
that particular m/z range will be degraded relative to the per-
formance outside of that range [50]. Similar statements can be
made with respect to the TIMS device. In the present work, we
emphasize the analytically useful charge capacity of the TIMS ana-
lyzer.
If one considers a line of charge trapped in a TIMS analyzer, an
associated electric field strength can be estimated based on funda-
mental principles. Consider that the rising edge of the EFG profile
(where the ions are stored according to their mobility) is ∼23 mm
long and the plateau region (over which the ions must “elute”) is
∼18 mm in length [46]. As an approximation, we therefore assume
106 charges, stored with uniform density in a line 23 mm in length,
 J.A. Silveira et al. / International Journal of Mass Spectrometry 413 (2017) 168–175 171

in free space (i.e., neglecting electrodes). The electric field strength

at radial distance, r, on the bisector of the line of charge is given by:
q 1
Er =  (4)
2εo r l2 + 4r 2
where q is the total charge on the line and l is the length of the line
of charges [51]. From Eq. (4), the electric field strength 2 mm from
the axis in the TIMS analyzer holding a million charges would be
roughly 0.6 V/cm—roughly two orders of magnitude less than the
TIMS analytical field strength.
It is useful to point out that as the ions elute from the TIMS
analyzer, they must traverse the plateau region of the EFG profile.
As the ions cross the plateau, they are spatially separated from the
stored ions and the influence of the stored ions on the eluting ions
is therefore diminished. Assuming the same conditions as given
above, the electric field on the EFG plateau due to the charges stored
on the rising is given by:
q
1 1

Ez = − (5)
4εo l lo l + lo
where lo is the distance from the near end of the line of stored
charges to the position on the plateau. By Eq. (5), the electric
field strength at a position 6 mm onto the plateau resulting from
a million charges stored on the rising edge would be roughly
0.09 V/cm—again, roughly two orders of magnitude less than the
TIMS analytical field strength.
These two field strength estimates suggest that the storage of
106 to 107 charges should be possible without having a major
impact on the analytical performance of the TIMS analyzer. Fur-
thermore, the estimates suggest that the radial field strength
experienced during storage will have more of an influence on the
ions than the axial field strength experience during elution. Thus,
one might expect it to be more likely that ions are lost radially
during storage rather than being shifted in their observed elution
voltage.
Fig. 2. Representative 2D TIMS-MS plot of tuning mix (a) and a BSA digest (b)
3.3. Performance evaluation of the original TIMS analyzer acquired on the 46 mm-long analyzer.

The charge capacity and performance of the original 46 mm-long

becomes nonlinear indicating that the capacity at the correspond-
tunnel was evaluated using tuning mix and tryptic peptides from
ing axial trapping position has been reached. Fig. 3c shows that a
a BSA digest (see Fig. 2). Fig. 2a reveals separation of the homol-
similar phenomenon is also observed for m/z 288 (2+) and m/z 1258
ogously substituted fluorinated triazatriphosphorine (tuning mix)
(2+). However, m/z 711 (2+) and m/z 942 (2+)—which reside in a
ions from other singly charged background ions along distinctive
particularly dense region of the 2D TIMS-MS spectrum—decrease
mobility/mass trendlines. Similarly, Fig. 2b demonstrates trendline
in abundance with additional accumulation time as a result of com-
dispersion of peptide ions that differ in their charge state (i.e., sepa-
petition for space in the trap by neighboring ions. Generally, as ion
ration of singly from multiply charged ions). The capacity of the trap
traps reach their capacity, loss of low m/z ions has been attributed
was determined from a plot of the measured ion intensities versus
to compression of the stable m/z range. Alternatively, loss of high
the accumulation time. Fig. 3a shows that as expected, the inten-
m/z ions often results from radial stratification effects [38]. That is,
sities for tuning mix ions increase linearly until the trap reaches
when the radius of a particular ion exceeds the radius of the con-
capacity whereupon the accumulation rate decreases. For tuning
fining potentials produced by the trap geometry, the ion will no
mix ions, the data indicate that this point occurs at ∼150 ms. Given
longer be stable and will be neutralized upon striking the surface of
that the sample produced 7 pA of continuous ion current (measured
an electrode [38,52]. In a TIMS experiment, ions of near-identical
downstream of the TIMS analyzer) and assuming a trapping effi-
mobility but dissimilar m/z are trapped in the same axial space;
ciency of 70% (see Section 3.2), the number of singly charged ions
thus, overfilling the trap can result in preferential loss of high m/z
stored is estimated to be ∼4.6 × 106 —in agreement with the first
ions that occupy the largest radii.
principles estimation of the total charge capacity discussed above.
A similar evaluation is more complex when tracking individual
ion intensities found in a tryptic BSA digest sample. At 1 pmol/␮L, 3.4. Performance evaluation of the parallel accumulation
the sample was found to produce a beam current roughly ∼20 pA measurement scheme
measured downstream of the TIMS analyzer. The relatively higher
beam current (roughly threefold greater compared to tuning mix) From the standpoint of maintaining a high duty cycle in a
and the presence of multiply charged ions together can, under serial analysis scheme, it is advantageous for the accumulation
some circumstances, produce effects that arise from mobility- time to be large with respect to the scan time. However, TIMS
based storage. Fig. 3b shows that singly charged ion intensities theory and experimental results have established that reduc-
largely increase upon extending the accumulation time up to ing the rate of the EFG scan improves
 the resolving power (i.e.,
200 ms, though the accumulation rate for m/z 791 (1+), for example, resolving power is proportional to 4 1/ˇ) [45,46]. Thus, reducing
172 J.A. Silveira et al. / International Journal of Mass Spectrometry 413 (2017) 168–175
Fig. 3. Tuning mix (a) and select singly (b) and doubly (c) charged ions from a
BSA digest plotted as a function of the accumulation time. Note that the data were
acquired on the 46 mm-long analyzer.
ˇ allows for improved separation performance, but does so at the
cost of a reduced duty cycle and/or mobility range. Past experi-
mental data demonstrate that employing a scan time of 50 ms, it
is feasible to achieve a resolving power exceeding 50 for a wide
range of singly charged ions [45]; assuming an accumulation time
of 150 ms, a duty cycle of ∼70% can be achieved. However, as shown
in Fig. 1c, a duty cycle of 100% is only asymptotically approached
as the accumulation time is extended further. Also, as discussed in
Section 3.1, the accumulation time has a practical limit set by the
capacity of the trap and the ion flux produced by the source/sample.
Thus, in practice, a serial measurement scheme cannot simul-
taneously yield a duty cycle of 100% and high TIMS resolving
power.
To overcome this principal limitation, we redesigned the tun-
nel region to incorporate an ion accumulation trap upstream of the
TIMS analyzer. Note that this second-generation spectrometer can
still be operated in the serial mode if so desired. In serial mode, one
may consider using the added length of tunnel for improved capac-
ity. That is, by simply extending the axial length of the rising edge
of the EFG profile, the charge capacity can be increased by a fac-
tor of three over the first-generation prototypes [53]. Though this
approach was previously successfully implemented, serial analysis
still suffers from the same principal duty cycle limitation outlined
above. Thus, as an alternative, the tunnel was segmented into two
regions configured with independent voltage control in a new mode
of operation.
As depicted in Fig. 1b, the new parallel accumulation mode
allows for the accumulation and analysis steps to occur simulta-
neously. Ions produced in the source are accumulated during the
analysis of a previously accumulated population, thus eliminating
the need to block ions using the deflector. Most notably, if the scan
time is equivalent to the accumulation time, the duty cycle can be
100% (see Fig. 1c). Note that in a serial analysis mode, the duty
cycle for equivalent operational parameters would only yield 50%.
Moreover, the new approach maintains the inherent flexibility to
trade off speed/duty cycle and/or mobility range when additional
resolving power is required.
A number of experiments were performed to explore the per-
formance of the new spectrometer. Fig. 4a contains a plot of the
resolving power as a function of the accumulation/analysis time
when operating TIMS at 100% duty cycle. The data indicate that as
expected from TIMS theory, the resolving power initially increases
upon extending the analysis time following a fourth-root depend-
ence. However, at analysis times exceeding 30 ms, the gain in
resolving power typically observed as a result of extending the scan
time appears to be largely offset by space charge effects. Though
most of the separation is achieved in a 30 ms scan, Fig. 4b shows that
the trap still has the capacity to store additional ions, evidenced by
the linear increase in the plot of the average peak area as a function
of accumulation time.
Next, to determine the extent of axial space charge broadening
that results from increasing the number of ions stored, the analysis
time was fixed at 120 ms while the duty cycle was varied by block-
ing ions entering the accumulation trap for a specified fraction of
the analysis sequence. In doing so, the average ion intensity across
the mass range was found to scale linearly with the duty cycle up to
100% (see Fig. 4c). At duty cycles between 1% and 10%, the resolv-
ing power was found to be fairly constant indicating that when the
number of ions stored is relatively low, broadening of the ion swarm
due to space charge effects is negligible (see Fig. 4d). However, upon
increasing the duty cycle (and thus the number of ions stored)
beyond 10%, a decrease in the resolving power (up to 30%) was
observed when the spectrometer was operated at 100% duty cycle.
Nevertheless, when operating at 100% duty cycle, the average TIMS
resolving power across the mobility range (R = 62) is still equiva-
lent to high resolution drift tube IMS separations [54]. Accordingly,
a representative two-dimensional TIMS-MS plot is shown in Fig. 4e
demonstrating separation comparable to serial mode of operation
(Fig. 2a).
It is important to note that 100% duty cycle was demonstrated
at an accumulation time of 120 ms and that because the accumula-
tion/analysis time is relatively long, charge capacity, for analytical
purposes, has been exceeded. In alternative timing methods, it is
possible to achieve a 100% duty cycle without effects due to charge
capacity. For example, limiting the accumulation/analysis time to
30 ms would achieve 100% duty cycle and yield similar resolving
power while avoiding charge capacity effects. Furthermore, higher
mobility resolution in combination with 100% duty cycle at a 30 ms
accumulation/analysis time can be achieved by limiting the mobil-
ity range analyzed. Limiting the mobility range results in a lower
scan rate, ˇ, which, as discussed previously, results in higher reso-
lution.
Finally, to assess the efficiency of storing and transferring ions
during TIMS, the relative signals produced in continuous and
trapping modes of operation were compared. Fig. 5a shows the
efficiency at 100% duty cycle for various accumulation/analysis
times plotted as a function of m/z. The data reveal no statistically
 J.A. Silveira et al. / International Journal of Mass Spectrometry 413 (2017) 168–175 173

Fig. 4. (a) Average resolving power for tuning mix ions (m/z 622–2722) versus accumulation/analysis time for operation of the 96 mm-long tunnel at 100% duty cycle. The
dotted line represents an extrapolation of the separation performance expected based on fourth-root dependence of the data acquired at relatively short accumulation/analysis
times (≤13 ms). The average TIMS peak area versus accumulation/analysis time is shown in (b). In (c) and (d), the duty cycle was varied using a fixed analysis time of 120 ms to
study the signal intensity (c) and the resolving power (d). Panel (e) contains a 2D TIMS-MS plot acquired on the 96 mm-long tunnel using the parallel accumulation/analysis
scheme operating at 100% duty cycle (120 ms accumulation/analysis time).

significant dependence when the experiment was performed at varied timescales indicated that (1) at 85 ms, the total charge capac-
accumulation/analysis times between 20 and 85 ms. Rather, the ity of the trap has not yet been reached and (2) that m/z-dependent
average trapping efficiency was found to be ∼70% for all timescales stability in the trap is not the primary factor that contributes to
investigated. That the efficiency remains constant for each m/z at ion losses. Rather, the m/z dependence observed in Fig. 5a is likely
174 J.A. Silveira et al. / International Journal of Mass Spectrometry 413 (2017) 168–175
Fig. 5. TIMS efficiency (a) and enhancement factor (b) at 100% duty cycle for various
accumulation/analysis times between 20 and 85 ms plotted as a function of m/z. In
(c), the signal observed for m/z 1222 in parallel accumulation mode is normalized
to the signal observed in MS-only (transmission) mode.
the result of differences in ion transmission injecting ions into and
capturing ions out of the TIMS tunnel.
In contrast to MS-only operation, ion packets are temporally dis-
persed based upon their mobility during TIMS. As a consequence,
operating at very short accumulation/analysis times (<20 ms) can
produce ion packets with narrow temporal peak widths that may
not overlap with the TOF acceleration pulse. Alternatively, at accu-
mulation/analysis times >20 ms, the average peak widths were
determined to be ∼400 ␮s such that more than four TOF spectra
were collected across the width of the TIMS peak. For accumu-
lation/analysis times between 20 and 85 ms, the ion intensities
measured at the centroid of peak were found to greatly exceed the
intensity for the same m/z ratio in the continuous beam. This effect
is demonstrated clearly in Fig. 5c wherein the signal for m/z 1222 is
measured with and without TIMS analysis. The transmission mode
signal is shown over a period of ∼47 ms and is normalized to an
intensity of one. The additional traces show the peak for m/z 1222
in parallel accumulation mode in an identical 47 ms time period.
It is clear that all the ions that would have otherwise appeared
as a continuous, low level signal now appear in a relatively high
intensity peak for a short duration. When operating at 100% duty
cycle, the apex of the mobility peak has a signal intensity 80 times
greater than that observed in transmission mode. The ratio of these
signals, termed the enhancement factor (see Fig. 5b), was found to
be between a factor of 60 and 120.
It is worth mentioning that it is because the ions are retained
and transmitted with relatively high efficiency, because a high
duty cycle is achieved, and because the IMS peaks are temporally
narrow, the signal intensities during ion elution are substantially
higher than in transmission mode. Importantly, assuming a con-
stant level of noise, this implies that the signal-to-noise ratio is
also concomitantly increased by the same factor. Furthermore, as
TIMS yields a mobility-based separation, we can also expect a
reduction in chemical background, further improving the signal-to-
noise ratio relative to the MS-only operational mode. Also, because
ions are eluted over time, a duty cycle enhancement in down-
stream devices can also be achieved, as recently described by Mann
et al. in a method termed “Parallel Accumulation SErial Fragmen-
tation” (PASEF) [31]. While space charge effects observed herein
were found to limit further gains in the resolving power for scan
times exceeding 30 ms, the capacity of the trap was found to effi-
ciently store additional ions without loss in performance. This
ability should prove useful in PASEF methods such that additional
precursor ions can be selected from a complex mixture in a single
experiment.
4. Conclusions
A new trapped ion mobility spectrometry (TIMS) apparatus has
been developed and evaluated. In addition to a TIMS analyzer, the
new spectrometer includes an upstream ion accumulation trap. The
new trap can be used to accumulate a group of ions while a pre-
viously accumulated group of ions is simultaneously analyzed. By
accumulating and analyzing ions in parallel rather than in series, a
duty cycle of 100% was demonstrated. The spectrometer was shown
to simultaneously trap a broad range of ions (m/z 622–2722) with
an average efficiency of ∼70%. Trapping efficiencies were found to
be constant over a range of accumulation/analysis times from 20 up
to 85 ms where an average resolving power of ∼60 was achieved. It
was found that, as currently constructed, the TIMS trap has an ana-
lytically useful charge capacity of ∼5 × 106 charges. The capacity
and ion beam current together set a limit on the accumulation time.
Compared to MS-only operation, the combination of high mobil-
ity resolving power, high ion utilization efficiency, and relatively
fast scan times achieved with TIMS is shown to improve the mass
spectral peak intensities by an average factor of ∼80.
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